fibrin and Periodontal-Diseases

Topics: Alveolar Bone Loss; Amino Acid Sequence; Bacteroides; Blood Coagulation Factors; Fibrin; Fibrinogen; Gingival Hemorrhage; Gingivitis; Humans; Molecular Sequence Data; Osteoblasts; Periodontal Diseases; Periodontitis; Thrombin; Tissue Plasminogen Activator

Regeneration of mineralized and soft connective tissue components of the attachment apparatus is the main goal in the treatment of periodontal diseases. Often, apical migration of epithelium (long junctional epithelium) effectively prevents the formation of bone and connective tissue attachment after periodontal surgery. The purpose of the present study was to compare conventional periodontal surgery combined with carbon dioxide laser and conventional periodontal surgery alone with respect to epithelial elimination and degree of necrosis of mucoperiosteal flaps. After signing a consent form, five patients with at least two comparable bilateral periodontal defects needing pocket elimination surgery participated in this study. The investigators randomly divided each side into test and control sites. Each patient received oral hygiene instruction and initial therapy prior to surgery. At surgery, the test site received a sulcular incision and carbon dioxide laser de-epithelialization of the outer and inner aspects of the flap. The control group received reverse bevel incision only. The surgeon performed open flap debridement on all teeth. At the time of surgery, the surgeon did a biopsy of each site and submitted specimens for histologic evaluation. A matched pairs t-test was used to analyze the data. The results show significant differences between the carbon dioxide laser and reverse bevel incision with respect to sulcular (P < or = 0.025) and gingival (external) (P < or = 0.01) flap surface epithelial elimination and tissue necrosis (P < or = 0.005). These results should be replicated with a larger number of subjects. The carbon dioxide laser eliminated sulcular and gingival (external) epithelium without disturbing underlying connective tissue. This finding supports the concept that the carbon dioxide wavelength has little or no effect on tissues beyond the target. However, neither laser nor blade eliminated all the epithelium. Researchers observed chronic inflammation in the control and test sites, with a predominance of plasma cells. Lining the sulcular and gingival (external) lased areas, investigators found coagulation necrosis covered by fibrin and coagulated blood. The laser appears to effectively remove epithelium at the time of surgery; however, future long-term, well-controlled quantitative histologic studies are needed to evaluate the effect of repeated carbon dioxide laser de-epithelialization of the gingival (external) surface of mucoperiosteal

Topics: Biopsy; Blood Coagulation; Carbon Dioxide; Cell Movement; Connective Tissue; Debridement; Epithelial Attachment; Epithelium; Female; Fibrin; Gingivectomy; Humans; Inflammation; Laser Therapy; Male; Matched-Pair Analysis; Necrosis; Oral Hygiene; Periodontal Diseases; Periodontal Pocket; Periodontium; Plasma Cells; Regeneration; Surgical Flaps; Wound Healing

Various studies have described the biological properties of the Leucocyte- and Platelet Rich Fibrin (L-PRF) such as the antimicrobial effect against wound bacteria, but less is known about the effect against periodontal pathogens. The aim of this study was to evaluate the antibacterial properties of the L-PRF membrane and L-PRF exudate against the main periopathogens cultured on agar plates and in planktonic solution. This study demonstrated the antibacterial effect of the L-PRF membrane against P. intermedia, F. nucleatum, and A. actinomycetemcomitans, but especially against P. gingivalis. The L-PRF exudate also showed a strong inhibition against P. gingivalis on agar plates. No inhibition could be observed for the other bacterial strains. Moreover, L-PRF exudate decreased the number of viable P.gingivalis in a planktonic solution in a dose-dependent way. However, A. actinomycetemcomitans showed an increased growth in planktonic solution when in contact with the L-PRF exudate.

Topics: Adult; Agar; Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents; Chlorhexidine; Female; Fibrin; Fusobacterium nucleatum; Humans; Leukocytes; Male; Microbial Sensitivity Tests; Middle Aged; Periodontal Diseases; Periodontium; Platelet-Rich Fibrin; Polymerase Chain Reaction; Porphyromonas gingivalis; Prevotella intermedia

Platelet-rich fibrin (PRF) has a controlled release of growth factors due to the fibrin matrix structure. Different centrifugation protocols were suggested for PRF preparation. Since the derivation method of PRF can alter its contents, in the present study it is aimed to investigate the cell contents and transforming growth factor beta-1 (TGF-β1), platelet-derived growth factor (PDGF-AB), vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-1 and-8 release from experimental PRF-type membranes obtained with different centrifugation times at 400 gravity. Three blood samples were collected from 20 healthy non-smoker volunteers. One tube was used for whole blood analyses. The other two tubes were centrifuged at 400 g for 10 minutes (group A) or 12 minutes (group B). Each experimental PRF-type membrane was placed in Dulbecco's Modified Eagle's Medium (DMEM)and at 1, 24 and 72 hours, TGF-β1, PDGF-AB, VEGF, MMP-1 and -8 release amounts were analysed by enzyme-linked immunosorbent assay (ELISA). The blood cell count of membranes was determined by subtracting plasma supernatant and red blood cell (RBC) mixture from the whole blood cell counts. At 72 hours, the VEGF level of group B was statistically higher than that of group A (p = 0.040). The centrifugation time was not found to influence the release of other growth factors, enzymes and cell counts. Within the limits of the present study, it might be suggested that centrifugation time at a constant gravity has a significant effect on the VEGF levels released from experimental PRF-type membrane. It can be concluded that due to the importance of VEGF in the tissue healing process, membranes obtained at 12-minute centrifugation time may show a superior potential in wound healing.

Topics: Adult; Biomarkers; Blood Cell Count; Blood Platelets; Case-Control Studies; Centrifugation; Female; Fibrin; Humans; Intercellular Signaling Peptides and Proteins; Male; Matrix Metalloproteinases; Periodontal Diseases; Platelet-Rich Plasma; Young Adult

In addition to releasing a pool of growth factors during activation, platelets have many features that indicate their role in the anti-infective host defense. The antimicrobial activities of platelet-rich plasma (PRP) and related plasma preparations against periodontal disease-associated bacteria were evaluated.. Four distinct plasma fractions were extracted in the formulation used commonly in dentistry and were tested for their antibacterial properties against three periodontal bacteria: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Fusobacterium nucleatum. The minimum inhibitory concentration of each plasma preparation was determined, and in vitro time-kill assays were used to detect their abilities to inhibit bacterial growth. Bacterial adhesion interference and the susceptibility of bacterial adherence by these plasma preparations were also conducted.. All plasma preparations can inhibit bacterial growth, with PRP showing the superior activity. Bacterial growth inhibition by PRP occurred in the first 24 hours after application in the time-kill assay. PRP interfered with P. gingivalis and A. actinomycetemcomitans attachment and enhanced exfoliation of attached P. gingivalis but had no influences on F. nucleatum bacterial adherence.. PRP expressed antibacterial properties, which may be attributed to platelets possessing additional antimicrobial molecules. The application of PRP on periodontal surgical sites is advisable because of its regenerative potential and its antibacterial effects.

Topics: Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents; Bacterial Adhesion; Bacterial Load; Bacteriological Techniques; Fibrin; Fusobacterium nucleatum; Humans; In Vitro Techniques; Male; Microbial Sensitivity Tests; Microbial Viability; Middle Aged; Periodontal Diseases; Plasma; Platelet Count; Platelet-Rich Plasma; Porphyromonas gingivalis; Time Factors

The objective of this research was to elucidate early events in periodontal wound healing/regeneration using histological and immunohistochemical techniques.. Routine critical-size, supraalveolar, periodontal defects including a space-providing titanium mesh device were created in 12 dogs. Six animals received additional autologous blood into the defect prior to wound closure. One animal from each group was killed for analysis at 2, 5, 9, 14 days, and at 4 and 8 weeks.. Both groups behaved similarly. Periodontal wound healing/regeneration progressed through three temporal phases. Early phase (2-5 days): heterogeneous clot consolidation and cell activation in the periodontal ligament (PDL) and trabecular bone was associated with PDL regeneration and formation of a pre-osteoblast population. Intermediate phase (9-14 days): cell proliferation (shown by PCNA immunostaining)/migration led to osteoid/bone, PDL and cementum formation. Late phase (4-8 weeks): primarily characterized by tissue remodelling/maturation. Fibrous connective tissue from the gingival mucosa entered the wound early, competing with regeneration. By day 14, the wound space was largely filled with regenerative and reparative tissues.. Activation of cellular regenerative events in periodontal wound healing/regeneration is rapid; the general framework for tissue formation is broadly outlined within 14 days. Most bone formation apparently originates from endosteally derived pre-osteoblasts; the PDL possibly acting as a supplementary source, with a primary function likely being regulatory/homeostatic. Blood accumulation at the surgical site warrants exploration; supplementation may be beneficial.

Topics: Alveolar Process; Animals; Blood; Blood Coagulation; Bone Matrix; Cell Differentiation; Cell Movement; Cell Proliferation; Cementogenesis; Collagen; Coloring Agents; Connective Tissue; Dental Cementum; Disease Models, Animal; Dogs; Erythrocytes; Fibrin; Fibroblasts; Gingiva; Immunohistochemistry; Osteoblasts; Osteogenesis; Periodontal Diseases; Periodontal Ligament; Regeneration; Time Factors; Wound Healing

Periodontitis involves bacterial infection, inflammation of the periodontium, degradation of gum tissue, and alveolar bone resorption, which eventually leads to loss of teeth. To study the role of the broad-spectrum protease plasmin in periodontitis, we examined the oral health of plasminogen (Plg)-deficient mice. In wild-type mice, the periodontium was unaffected at all time points studied; in Plg-deficient mice, periodontitis progressed rapidly, within 20 weeks. Morphological study results of Plg-deficient mice revealed detachment of gingival tissue, resorption of the cementum layer, formation of necrotic tissue, and severe alveolar bone degradation. IHC staining showed massive infiltration of neutrophils in the periodontal tissues. Interestingly, doubly deficient mice, lacking both tissue- and urokinase-type plasminogen activators, developed periodontal disease similar to that in Plg-deficient mice; however, mice lacking only tissue- or urokinase-type plasminogen activator remained healthy. Supplementation by injection of Plg-deficient mice with human plasminogen for 10 days led to necrotic tissue absorption, inflammation subsidence, and full regeneration of gum tissues. Notably, there was also partial regrowth of degraded alveolar bone. Taken together, our results show that plasminogen is essential for the maintenance of a healthy periodontium and plays an important role in combating the spontaneous development of chronic periodontitis. Moreover, reversal to healthy status after supplementation of Plg-deficient mice with plasminogen suggests the possibility of using plasminogen for therapy of periodontal diseases.

Topics: Alkaline Phosphatase; Animals; Disease Models, Animal; Fibrin; Fibrinolysin; Immunohistochemistry; Inflammation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Models, Genetic; Neutrophils; Periodontal Diseases; Periodontitis; Time Factors

Infectious process frequently results in extensive bone resorption and defect, periradicular or periapical lesions, or vertical fracture with infected sinus tract. When tooth extraction is mandated it typically results in additional bone loss in the buccal or lingual cortical plate. Immediate guided bone regeneration (GBR) and implant fixation at an infected site is frequently complicated by soft-tissue dehiscence, membrane exposure, and implant failure. The objective of this research is to assess the feasibility of immediate bone augmentation (IBA) after purulent tooth extraction, employing a dedicated titanium membrane. An intrasulcular incision was made around the tooth to be extracted and extended to 2 adjacent teeth while maintaining the papillae. Vertical releasing incisions were made to mobilize the mucoperiosteal flap. Cautious tooth extraction was executed utilizing conventional measures and was followed by meticulous curettage of the infected and granulated tissue in the socket. Titanium membranes were applied to the socket walls followed by socket filling with autologous platelet-rich fibrin and primary closure. Eight or more weeks later membrane removal and implant placement were performed. Of the 15 patients who underwent this procedure, 7 patients (47%) had early membrane exposure (between weeks 2 and 6), which was treated conservatively. No infection or early membrane removal was reported. All patients achieved sufficient bone augmentation, and 8 patients received implants without any additional GBR. IBA after infected tooth extraction, using titanium membrane application was feasible and safe and yielded adequate bone filling to support implant fixation at > or =8 weeks. Further studies need to evaluate if the titanium membrane helped in any way to inhibit plaque accumulation or resist infection in cases of early membrane exposure.

Topics: Adult; Bacterial Infections; Biocompatible Materials; Bone Regeneration; Curettage; Dental Fistula; Dental Implants; Feasibility Studies; Female; Fibrin; Granulation Tissue; Guided Tissue Regeneration, Periodontal; Humans; Male; Membranes, Artificial; Middle Aged; Periapical Diseases; Periodontal Diseases; Platelet-Rich Plasma; Titanium; Tooth Diseases; Tooth Extraction; Tooth Socket; Treatment Outcome

Cell-surface proteoglycans participate in several biological functions including interactions with adhesion molecules, growth factors and a variety of other effector molecules. Accordingly, these molecules play a central role in various aspects of cell-cell and cell-matrix interactions. To investigate the expression and distribution of the cell surface proteoglycans, syndecan-1 and -2, during periodontal wound healing, immunohistochemical analyses were carried out using monoclonal antibodies against syndecan-1, or -2 core proteins. Both syndecan-1 and -2 were expressed and distributed differentially at various stages of early inflammatory cell infiltration, granulation tissue formation, and tissue remodeling in periodontal wound healing. Expression of syndecan-1 was noted in inflammatory cells within and around the fibrin clots during the earliest stages of inflammatory cells infiltration. During granulation tissue formation it was noted in fibroblast-like cells and newly formed blood vessels. Syndecan-1 was not seen in newly formed bone or cementum matrix at any of the time periods studied. Syndecan-1 expression was generally less during the late stages of wound healing but was markedly expressed in cells that were close to the repairing junctional epithelium. In contrast, syndecan-2 expression and distribution was not evident at the early stages of inflammatory cell infiltration. During the formation of granulation tissue and subsequent tissue remodeling, syndecan-2 was expressed extracellularly in the newly formed fibrils which were oriented toward the root surface. Syndecan-2 was found to be significantly expressed on cells that were close to the root surface and within the matrix of repaired cementum covering root dentin as well as at the alveolar bone edge. These findings indicate that syndecan-1 and -2 may have distinctive functions during wound healing of the periodontium. The appearance of syndecan-1 may involve both cell-cell and cell-matrix interactions, while syndecan-2 showed a predilection to associate with cell-matrix interactions during hard tissue formation.

Topics: Alveolar Process; Animals; Antibodies, Monoclonal; Blood Coagulation; Blood Vessels; Cell Communication; Dental Cementum; Disease Models, Animal; Epithelial Attachment; Extracellular Matrix; Extracellular Space; Fibrin; Fibroblasts; Gene Expression; Granulation Tissue; Immunoenzyme Techniques; Immunohistochemistry; Male; Membrane Glycoproteins; Neovascularization, Physiologic; Periodontal Diseases; Periodontal Ligament; Proteoglycans; Rats; Rats, Inbred Lew; Syndecan-1; Syndecan-2; Syndecans; Tooth Root; Wound Healing

The plasminogen activator system has been proposed to play a role in proteolytic degradation of extracellular matrices in tissue remodeling, including wound healing. The aim of this study was to elucidate the presence of components of the plasminogen activator system during different stages of periodontal wound healing.. Periodontal wounds were created around the molars of adult rats and healing was followed for 28 days. Immunohistochemical analyses of the healing tissues and an analysis of the periodontal wound healing fluid by ELISA were carried out for the detection of tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA), and 2 plasminogen activator inhibitors (PAI-1 and PAI-2).. During the early stages (days 1 to 3) of periodontal wound healing, PAI-1 and PAI-2 were found to be closely associated with the deposition of a fibrin clot in the gingival sulcus. These components were strongly associated with the infiltrating inflammatory cells around the fibrin clot. During days 3 to 7, u-PA, PAI-1, and PAI-2 were associated with cells (particularly monocytes/macrophages, fibroblasts, and endothelial cells) in the newly formed granulation tissue. During days 7 to 14, a new attachment apparatus was formed during which PAI-1, PAI-2, and u-PA were localized in both periodontal ligament fibroblasts (PDL) and epithelial cells at sites where these cells were attaching to the root surface. In the periodontal wound healing fluid, the concentration for t-PA increased and peaked during the first week. PAI-2 had a similar expression to t-PA, but at a lower level over the entire wound-healing period.. These findings indicate that the plasminogen activator system is involved in the entire process of periodontal wound healing, in particular with the formation of fibrin matrix on the root surface and its replacement by granulation tissue, as well as the subsequent formation of the attachment of soft tissue to the root surface during the later stages of wound repair.

Topics: Animals; Blood Coagulation; Cell Adhesion; Disease Models, Animal; Endothelium, Vascular; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Fibrin; Fibroblasts; Follow-Up Studies; Gingival Crevicular Fluid; Granulation Tissue; Immunohistochemistry; Macrophages; Male; Monocytes; Periodontal Diseases; Periodontal Ligament; Plasminogen Activator Inhibitor 1; Plasminogen Activator Inhibitor 2; Plasminogen Activators; Rats; Rats, Inbred Lew; Serine Proteinase Inhibitors; Tissue Plasminogen Activator; Tooth Root; Urokinase-Type Plasminogen Activator; Wound Healing

Studies on periodontal repair to denuded root surfaces have suggested that initial clot adhesion to the root surface may be important for the nature of subsequent healing. To study this hypothesis, circumferential periodontal defects, approximately 5 mm in vertical dimensions, were surgically created and immediately treated around the mandibular premolars in 4 beagle dogs. Prior to wound closure, the root surfaces were treated with either the anticoagulant heparin or with saline. Tissue blocks were obtained at sacrifice 4 weeks after surgery. Histometric analysis showed that connective tissue repair to the root surface averaged 50% of the defect height for heparin-treated teeth as compared to 95% for saline-treated teeth. Junctional epithelium amounted to an average of 33% of the defect height in heparin-treated teeth in contrast to 5% following saline treatment. It can be concluded that heparin treatment of the root surface compromises connective tissue repair, confirming clot adhesion as one prerequisite for connective tissue repair of periodontal defects.

Topics: Alveolar Bone Loss; Alveolar Process; Animals; Blood Coagulation; Bone Regeneration; Connective Tissue; Dental Cementum; Dogs; Epithelium; Fibrin; Heparin; Male; Periodontal Diseases; Sodium Chloride; Tooth Root; Wound Healing

Crevicular fluid (CF) samples were collected by paper strips from healthy and diseased sites. The molecular distribution of fibrin and fibronectin in CF and plasma samples was investigated using SDS-polyacrylamide gel electrophoresis and specific immunoglobulins. Intact fibrin was found in all CF samples. In addition several bands with both lower and higher molecular weights than intact fibrin were seen. These bands were not present in the plasma samples. The low molecular weight bands are suggested to represent degradation products of fibrin. The high molecular bands could be fibrin-fibronectin complexes or fragments of large fibrin polymers. Fibronectin degradation products, but not intact fibronectin, were seen in both healthy and diseased samples.

Topics: Blood; Electrophoresis, Polyacrylamide Gel; Fibrin; Fibrin Fibrinogen Degradation Products; Fibrinogen; Fibronectins; Gingival Crevicular Fluid; Humans; Immunoblotting; Periodontal Diseases; Periodontium; Sodium Dodecyl Sulfate

This study presents the morphological sequence of events in initial blood clot formation on various root surfaces of freshly extracted human teeth. Four teeth with periodontal disease (PD) and three teeth without PD were extracted and the roots sectioned into halves. Those root surfaces with PD had four treatment areas: (1) intact periodontal ligament (PDL), (2) unplanned PD, (3) PD plus planed, (4) PD plus planed plus application of pH 1 citric acid (CA). The roots with no PD had three treatment areas: (1) intact PDL, (2) planed, (3) planed plus CA. Both root halves were reinserted together into the original extraction site. Each root half was then removed at either zero, one, two or four minutes and prepared for scanning electron microscope (SEM) evaluation. SEM observations suggested that plasma proteins were deposited initially on all root surfaces. Platelets and erythrocytes enmeshed in fibrin deposited most rapidly and consistently over the plasma protein layer where intact PDL was present. Similar observations were noted on the planed plus CA surfaces and appeared to occur at an earlier time than on the planed-only surfaces. A constant feature at all time periods was the absence of organized clot formation over the plaque-free zone of the PD root surfaces. By two minutes all surfaces, except the plaque-free zone of the PD area, appeared to have clot lysis occurring. While clot formation appeared to occur more rapidly over surfaces in the non-PD roots, no marked morphological differences in clot formation were observed between PD and non-PD teeth with similar root surface treatments.

Topics: Blood Coagulation; Blood Platelets; Blood Proteins; Citrates; Citric Acid; Dental Prophylaxis; Dental Scaling; Dentin; Erythrocytes; Fibrin; Humans; Microscopy, Electron, Scanning; Periodontal Diseases; Periodontal Ligament; Tooth Root

The purpose of the study was two-fold: to determine the nature of stainable deposits on periodontally diseased root surfaces subsequent to in vivo scaling and root planing procedures, and to quantify the distribution of residual plaque on instrumented root surfaces. Thirty molar and 30 nonmolar teeth which were condemned for periodontal or prosthetic reasons and had proximal probing depths of 4 to 7 mm were treated. Half of these were instrumented with I.U. curettes and the other half with an ultrasonic scaling device. Instrumentation was continued until the root surface felt hard and smooth to an explorer tip. The location of the gingival margin was recorded by notching the treated proximal surface with a No. 1/2 round bur. Twenty control teeth, 10 molar and 10 nonmolar, were extracted without instrumentation. Control and experimental teeth were irrigated with saline and stored in a 2.5% glutaraldehyde fixative solution until the time of assessment. All teeth were stained with a 0.5% solution of toluidine blue, and the amount of residual stained material and calculus was assessed under the stereomicroscope using an eyepiece fitted with a 10 X 10 optical grid. Stained deposits were marked by placing small V-shaped notches in the adjacent root surface as an aid to identification after the specimens were processed for scanning electron microscopic (SEM) examination. The nature of stained deposits on selected teeth was then characterized using the SEM. Treated root surfaces were also surveyed in detail to assess the quantity and extent of residual plaque deposits. The findings showed that although a large percentage of the treated proximal root surface may possess stainable deposits, these surfaces were often unexpectedly free of microbial organisms. In this study, the majority of stained deposits were composed of adherent fibrin and instrumentation debris. When bacterial plaque was present, it was usually found in small "mini-colonies" smaller than 0.5 mm across. Such findings cast doubt on the validity of using histologic and disclosing stains as an indicator for the presence of bacterial plaque immediately after instrumentation. Although only partially effective in removing subgingival calculus, both methods of instrumentation in this study appeared to be remarkably effective in bacterial debridement of subgingival root surfaces.

Topics: Bacteria; Dental Calculus; Dental Plaque; Dental Prophylaxis; Dental Scaling; Fibrin; Humans; Microscopy, Electron, Scanning; Periodontal Diseases; Tooth Root; Ultrasonic Therapy

Topics: Animals; Blood Coagulation; Fibrin; Male; Periodontal Diseases; Rats; Rats, Inbred Strains; Wound Healing

Topics: Adult; Alveolar Process; Bone Transplantation; Fibrin; Humans; Middle Aged; Periodontal Diseases; Transplantation, Homologous

Topics: Alveolar Process; Alveolectomy; Animals; Bone Regeneration; Bone Resorption; Cementogenesis; Dogs; Epithelium; Fibrin; Gingiva; Periodontal Diseases; Subgingival Curettage; Wound Healing